The effect of polyamines of DNA synthesis in vitro

Nuclei and DNA-dependent polymerases alpha and beta were isolated from exponentially growing baby hamster kidney 21/C13 cells and were used to study the effects of polyamines on DNA synthesis in vitro. The greatest effect was observed with spermine, which inhibited both nuclear DNA synthesis and the activity of partially purified DNA polymerase alpha. At 2.5 mM spermine, the maximum concentration used, we observed 58 and 68% inhibition of DNA synthesis by isolated nuclei and polymerase alpha, respectively. In contrast, spermidine caused a small increase in nuclear DNA synthesis at low concentrations (0.5 mM) and inhibition at higher concentrations (2.5 mM); it had no significant effect on the partially purified polymerase alpha. Neither polyamine had any appreciable effect on polymerase beta activity. The results are consistent with the concept that DNA polymerase alpha catalyses the observed DNA synthesis in isolated nuclei.